Project Summary/Abstract Despite the prevalence and large mortality rate associated with colon cancer, clinicians are still in search for suitable drugs to treat the disease. More recently, efforts to treat cancer have focused on the use of therapeutics that target epigenetic mechanisms to correct gene regulatory programs that have gone awry during neoplastic transformations. These ?epigenetic drugs? have had success in clinical trials for the treatment of blood cancers, however, their use in the treatment of solid tumor malignancies has been less effective. Many explanations on why epigenetic drugs do not seem to work on solid tumors have been proposed, including: drug stability or diffusion limitations, development of post-chemo resistance, and patient-specific genetic differences. Mechanical heterogeneity within solid tumor tissues, and the potential detrimental impact that is could have on cancer cell epigenetics, has not thoroughly been considered. Our proposal aims to build novel tools and technologies that enable spatial epigenetic and mechanical measurements in tumor tissues to help reveal these molecular connections for the first time. We also aim to develop a novel epigenetic inhibitor capable of targeting DNA methylation with locus-specific precision ultimately for the treatment of colon cancer but with implications beyond. Furthermore, outside of tumor tissue mechanics, spatial epigenomics will allow us to determine how the cancer cell epigenome responds to the complex drug profiles created when trying to deliver therapeutics (especially in combination) into solid tumor tissues. These studies will reveal how the mechanical heterogeneity found within colon cancer tissues contribute to heterogeneity in the epigenetic landscape of cancer cells to influence tumor prognosis and the efficacy of epigenetic drugs.